Extracellular Vesicle RNA Sequencing Reveals Dramatic Transcriptomic Alterations Between Metastatic and Primary Osteosarcoma in a Liquid Biopsy Approach
Extracellular Vesicle RNA Sequencing Reveals Dramatic Transcriptomic Alterations Between Metastatic and Primary Osteosarcoma in a Liquid Biopsy Approach
复制标题
细胞外囊泡 RNA 测序揭示液体活检方法中转移性骨肉瘤和原发性骨肉瘤之间的巨大转录组变化
DOI：
10.1245/s10434-018-6642-z
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发表时间：
2018-09-01
影响因子：
3.7
通讯作者：
Zhang, Weibin
中科院分区：
文献类型：
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作者：
Bao, Qiyuan;Gong, Liangzhi;Zhang, Weibin
骨肉瘤（osteosarcoma，OS）是一种原发灶广泛切除后仍有高度转移的骨恶性肿瘤.仍然缺乏液体活检方法来检测残留病变和确定治疗靶点。在这份报告中，我们的目的是跟踪转移OS通过细胞外囊泡（EV）RNA谱在一个非侵入性manners.MethodsWe应用RNA测序10匹配转移和原发OS EV样本，包括两对细胞系和三对血浆，并比较表达的突变，基因表达，融合转录，和选择性剪接（AS）之间的转移性和原发性OS在转录组水平。额外配对的组织/EV进行测序和公共数据集被用来验证EV为基础的转移bipolis. ResultsEV的特点是通过大小分析，免疫标记，和形态学检查。与非转移性对应物相比，转移性OS中观察到突变负荷急剧增加。表达谱的层次聚类将转移性EV与非转移性EV区分开，其中特征富集于细胞粘附信号传导和酪氨酸激酶途径。此外，在EV RNA中鉴定了30种癌症相关基因融合，因为AS事件往往在转移性EV中更频繁地观察到。进一步的研究表明，超过70%的EV表达点突变可以在配对的细胞系/EV和组织/EV分析中得到验证，并且表达特征显著预测了来自公共dataset.ConclusionWe的42名患者的5年生存率，我们已经证明了一种基于液体活检的方法用于跟踪癌症转录组学改变，这是转移性OS的预后和治疗生物标志物的有希望的来源。
BackgroundOsteosarcoma (OS) is a highly metastasizing bone malignancy despite wide surgical resection of the primary lesion. A liquid biopsy approach to detect residual disease and identify therapeutic targets is still lacking. In this report, we aimed to track the metastasis of OS via extracellular vesicle (EV) RNA profiling in a non-invasive manner.MethodsWe applied RNA sequencing for 10 matched metastatic and primary OS EV samples, including two pairs of cell lines and three pairs of plasma, and compared the expressed mutation, gene expression, fusion transcript, and alternative splicing (AS) between metastatic and primary OS at the transcriptome-wide level. Additional paired tissue/EVs were sequenced and public datasets were used to validate the EV-based metastatic biopsy.ResultsEVs were characterized through size-profiling, immunolabeling, and morphological examination. A drastic increase of mutation burden was observed in metastatic OS versus the non-metastatic counterpart. Hierarchical clustering of the expression profiles differentiated the metastatic EVs from the non-metastatic, with a signature enriched in cell-adhesion signaling and tyrosine kinase pathways. Moreover, 30 cancer-related gene fusions were identified in EV RNA as AS events tend to be more frequently observed in metastatic EVs. Further investigation suggested that over 70% of expressed point mutations from EVs could be validated in paired cell line/EV and tissue/EV analyses, and the expression signature significantly predicted 5-year survivorship of 42 patients from a public dataset.ConclusionWe have demonstrated a liquid biopsy-based approach for tracking cancer transcriptomic alterations, which is a promising source of prognostic and therapeutic biomarkers for metastatic OS.Clinical Trial RegistrationNCT03108677.